CN109828365B - Mirau type super-resolution interference microscope objective - Google Patents
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- JRPBQTZRNDNNOP-UHFFFAOYSA-N barium titanate Chemical compound [Ba+2].[Ba+2].[O-][Ti]([O-])([O-])[O-] JRPBQTZRNDNNOP-UHFFFAOYSA-N 0.000 claims description 7
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
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- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 description 5
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Abstract
The invention discloses a Mirau type super-resolution interference microscope objective, which realizes sub-nanometer axial resolution by utilizing the interference microscope objective, breaks through diffraction limit by utilizing a microsphere lens and realizes two-dimensional transverse super-resolution. The super-resolution interference microscope objective lens is formed by fusing an interference microscope objective lens and a microsphere lens, the microsphere lens is added between the interference microscope objective lens and a sample to be detected, parameters of a reference flat plate, a light splitting flat plate and the microscope objective lens are optimally designed, clear three-dimensional super-resolution interference fringes are obtained, and three-dimensional high-resolution imaging is realized. The method can realize transverse super-resolution by a simple optical means, obtain the three-dimensional high-resolution information of the microstructure to be measured, does not need to carry out complex marking treatment on the sample, can realize rapid and nondestructive measurement, and has strong practical value.
Description
Technical Field
The invention belongs to the field of interference microscopic measurement, and particularly relates to a Mirau type super-resolution interference microscope objective.
Background
The conventional common micro-nano observation means mainly comprises the following steps according to an imaging mechanism: optical microscopes, scanning probe-type microscopes, and electron microscopes. Compared with other imaging modes, the optical microscope has the advantages of real time, no mark, no damage, simplicity, convenience, low cost, low maintenance cost and the like, and plays an irreplaceable role in the aspect of exploring the micro world by human beings. However, due to the diffraction limit, the optical microscope has a lateral resolution of half the wavelength, which is only 200 nm at the highest. The actual requirements of people for microstructure observation cannot be met. Wangmangbo et al, the university of Manchester, UK, utilize silica microspheres with a radius of 2-9 μm to improve the resolution of visible light micro-imaging to about 50 nm, and successfully utilize microspheres in combination with a microscope objective to realize two-dimensional super-resolution imaging (Nature Communications, 2011, 2: 218-0.).
The white light micro-interference technology utilizes the low coherence characteristic of broadband illumination light, obtains a plurality of interference images by driving the vertical scanning of an interference microscope objective, solves the position with zero optical path difference, realizes the three-dimensional measurement of surface micro-morphology, and has the axial resolution reaching the sub-nanometer level. The Mirau type interference microscope objective is the most commonly used interference microscope objective. The conventional Mirau type interference microscope objective consists of a microscope objective, a light splitting flat plate and a reference flat plate and has the advantage of compact structure. When the Mirau type interference microscope objective is designed, in the interference aspect, the light splitting flat plate and the reference flat plate are made of the same material and thickness (the typical material is fused quartz); in the aspect of two-dimensional imaging, a microscope objective, a light splitting flat plate and a reference flat plate are used for jointly correcting aberration. The lateral resolution of this technique is still limited by diffraction effects, up to only 200 nm. The method can improve the transverse resolution while keeping the sub-nano axial high resolution, and is very important for three-dimensional imaging.
The combination of the microsphere transverse super-resolution imaging technology and the interference microscopic axial high-resolution technology is an effective means for realizing three-dimensional high-resolution imaging. The Wangfei of Chinese academy of sciences adopts two modes to carry out three-dimensional high-resolution imaging: one method is that barium titanate microspheres are sown on the surface of a sample, water is injected, the microspheres are immersed in the water, a cover glass is covered, and then a Linnik type interference microscope objective is used to combine the microspheres for observation; another method is to directly spread polystyrene microspheres on the surface of the sample and use Linnik type interference microscope objective lens to combine the microspheres for observation (Scientific Reports, 2016, 6(1):24703, microlens-based three-dimensional super-resolution interferometer CN 201510309193.3). The Ivan Kassamakov team at Helsinki university of Finland used a Nikon 50-fold Mirau type interference objective lens in combination with microspheres to achieve three-dimensional imaging for Blu-ray discs (Scientific Reports, 2017, 7(1): 3683.).
The Linnik type interference microscope objective belongs to a light splitting path system, is not compact in structure, can only drive the micro displacement of a reference reflector in order to realize vertical scanning, and cannot be directly used on a white light microscope interferometer. In addition, optical imaging aberrations introduced by the microspheres are not eliminated. In addition, when the optical path matching device is used, the axial position of the reference reflector needs to be manually adjusted, and only the optical path matching of the reference optical path and the test optical path under the central wavelength is realized. Therefore, the structure can not directly use the vertical scanning structure of the white light micro-interferometer, the driving reference reflector needs to be controlled to carry out micro-displacement, the aberration is under-corrected, the optical path matching is difficult, a super-resolution interference micro-objective assembly is difficult to form, and the structure is not suitable for being directly used on a commercial white light micro-interferometer.
The Ivan Kassamakov team at Helsinki university of Finland directly uses a conventional Mirau type interference microscope objective lens to counteract the optical path difference introduced by the microsphere lens on the test arm by adjusting the distance between the beam splitting plate and the reference plate. The method is based on a conventional interference microscope objective, the adjusting range is less than hundred micrometers, and the optical path introduced by the large-size microspheres cannot be matched. The method is only suitable for the microspheres with small sizes, and the obtained imaging field of view is small. In addition, the problems of imaging aberration, chromatic dispersion and the like introduced by the microsphere lens are not corrected. The problems of small application range, large error of measurement results and the like exist.
The completed work is to directly spread the microspheres on the surface of a sample to be measured, and then to measure the three-dimensional appearance by combining the interference microscope objective with the microspheres. The spreading of the microspheres is random and not a standardized optical imaging procedure. And the immersion method has the problems of easy volatilization, short observation time and poor experiment repeatability. The preparation of the microsphere lens film has many advantages: by combining the barium titanate microspheres and the PDMS film, compared with other microspheres with low refractive index, the high-refractive-index PDMS film has higher magnification and imaging quality and can obtain a larger field of view; in the immersion mode, a solid immersion mode is adopted, so that the problem of liquid volatilization is avoided; the microsphere lens film can be directly placed on the surface of a sample, is simple and convenient to operate, can be directly carried and used on a white light micro-interferometer, and has high practical value.
Disclosure of Invention
The invention aims to provide a Mirau type super-resolution interference microobjective, which realizes sub-nanometer axial resolution by using the interference microobjective, breaks through diffraction limit by using a microsphere lens film and realizes two-dimensional transverse super resolution.
The technical solution for realizing the purpose of the invention is as follows: a Mirau type super-resolution interference microscope objective comprises a microscope objective, a reference flat plate, a light splitting flat plate and a micro-ball lens film, wherein the microscope objective, the reference flat plate, the light splitting flat plate and the micro-ball lens film are sequentially arranged along a common optical axis;
the illumination light sequentially passes through the microscope objective and the reference flat plate, and is divided into two beams at the light splitting plane of the light splitting flat plate, one beam is incident to the reference surface of the reference flat plate and returns to form reference light, and the other beam is incident to a sample to be detected and returns after passing through the microsphere lens film to form test light; the reference light and the test light are overlapped on the light splitting plate and then interfere with each other, and the reference light and the test light are emitted after passing through the reference plate and the microscope objective.
Compared with the prior art, the invention has the remarkable advantages that:
(1) the invention combines the microsphere lens film and the interference microscope objective, breaks the limit of the optical diffraction limit to the transverse resolution of the optical microscope, obtains height information by utilizing the principle of white light interference and realizes three-dimensional super-resolution imaging.
(2) Compared with a microsphere combined Linnik type interference microscope objective, the invention has a more compact structure. In the actual operation process, the reference arm and the test arm do not need to be adjusted, and the operation is simple. Errors generated by light splitting paths cannot be introduced, the influence of the environment is small, and the problem of poor interference resistance is solved. Compared with a Mirau type interference microscope objective with the Nikon 50 times, the position of the light splitting plate does not need to be adjusted. And the method is suitable for large-size microspheres, and can obtain a larger imaging field of view. Compared with the two modes, the invention solves the problems of additional spherical aberration, chromatic aberration and the like introduced by the microspheres, and has more accurate measurement result and smaller measurement error.
(3) Compared with other microspheres with low refractive index, the microsphere lens film prepared by combining the barium titanate microspheres and the PDMS has higher magnification and imaging quality, and obtains a larger imaging field of view. And moreover, by adopting a solid immersion mode, the super-resolution phenomenon of the high-refractive-index microspheres is obtained (the super-resolution phenomenon does not exist when the high-refractive-index microspheres are placed in the air), and compared with liquid immersion, the complex operation on a sample is not needed, the observation result cannot be changed along with the volatilization of the immersion liquid, and the measurement has repeatability. In the aspect of operation, the micro-sphere lens film is only required to be directly placed on the surface of a sample, the Mirau type interference microscope objective after optimized design is carried on the white light microscope interferometer, the three-dimensional result of the microstructure to be measured can be obtained according to the conventional operation mode of the white light microscope interferometer, and the limit of diffraction limit is broken. The interferometer has the advantages of simple operation, stable structure, repeated use and convenience for large-scale application on commercial interferometers.
(4) The method has no special requirements on the environment during imaging, does not need to carry out complex marking on a sample, and can realize rapid, non-invasive and lossless three-dimensional super-resolution imaging, thereby having wide application space in the field of nano imaging.
Drawings
FIG. 1 is a structural diagram of a Mirau type super-resolution interference microscope objective lens.
FIG. 2 is a parameter labeling diagram according to the present invention.
FIG. 3 is an interference image obtained by a white light micro-interferometer by a 50-time Mirau type super-resolution interference micro objective with optimized design.
FIG. 4 is a three-dimensional information map of the surface topography of the sample obtained from the interferogram of FIG. three.
FIG. 5 is a diagram of an apparatus for interferometry using a Mirau type super-resolution interference microscope objective of the present invention.
Detailed Description
The present invention is described in further detail below with reference to the attached drawing figures.
With reference to fig. 1 and 5, a Mirau type super-resolution interference microscope objective comprises a microscope objective 1, a reference flat plate 2, a light splitting flat plate 3 and a microsphere lens film 4 which are sequentially arranged along a common optical axis, wherein the microsphere lens film 4 is arranged on the top surface of a sample 5 to be tested, a Mirau type interference optical path structure is adopted, light is split by the light splitting flat plate 3, the reference flat plate 2 is positioned on a reference arm, and the microsphere lens film 4 is positioned on a test arm.
The illumination light 9 passes through the lens 11 and the spectroscope 10, then sequentially passes through the microscope objective 1 and the reference flat plate 2, is divided into two beams at the light splitting plane of the light splitting flat plate 3, one beam is incident to the reference surface of the reference flat plate 2 and returns to form reference light, and the other beam is incident to the sample to be tested 5 and returns after passing through the microsphere lens film 4 to form test light; the reference light and the test light are overlapped on the light splitting plate 3 and then interfere with each other, the reference light and the test light are emitted after passing through the reference plate 2 and the microscope objective 1, emergent light is received by the image acquisition system 8 after passing through the spectroscope 10 and the lens 11, and the microscopic characteristics of the microstructure to be tested are analyzed according to interference fringes acquired by the image acquisition system 8.
The illuminating light is a visible light broad spectrum, and aberration is corrected for the full spectrum of the working wavelength of the illuminating light source during design.
The interference microscope objective lens is formed by the microscope objective lens 1, the reference flat plate 2 and the light splitting flat plate 3.
In the reference flat plate 2, the surface close to the microscope objective 1 is a reference surface, namely an A surface; in the spectroscopic plate 3, the surface close to the reference plate 2 is a spectroscopic plane, i.e., a B-surface.
The magnification of the microscope objective 1 is not less than 10 times.
The microsphere lens film 4 is a film prepared by coating a layer of material on the surface of a microsphere with a high refractive index to immerse the microsphere solid.
At the center wavelength λ of the broad spectrum illumination light0The sum of the optical paths of the light splitting plate 3 and the microsphere lens film 4 is equal to the optical path of the reference plate 2.
The microsphere lens film 4 coats a layer of film on the surface of the microsphere array, and with reference to fig. 2, the microsphere array is equivalent to a microsphere flat plate 7, and the film is equivalent to a film flat plate 6. The refractive index of the film flat plate 6 is the refractive index of the film, and the thickness of the film flat plate 6 is the thickness of the microsphere lens film 4 minus the thickness of the microsphere flat plate 7; the refractive index of the microsphere flat plate 7 is the refractive index of the microsphere, and the thickness of the microsphere flat plate 7 is the diameter of the microsphere; at a central wavelength λ0Next, the following equation is satisfied:
n1t1+h1=n2t2+h2+n3t3+n4t4
wherein n is1For reference to the refractive index of the plate 1, n2Is the refractive index of the spectroscopic plate 2, t1For reference to the thickness of the plate 1, t2Is the thickness of the dispersing plate 2, h1For the spacing of the reference plate 1 and the spectroscopic plate 2, h2Is the interval between the light-splitting plate 2 and the film plate 6, t3Is the thickness of the film plate 6, n3Is the refractive index of the thin-film flat plate 6,t4is the diameter of the microspherical plate 7, n4The refractive index of the microsphere flat plate 7.
Although the thickness relationship of the reference plate 2, the light splitting plate 3, the film plate 6 and the microsphere plate 7 is optimally controlled, the center wavelength lambda of the reference arm and the test arm is realized0Lower aplanatic. However, since the flat plate is made of different materials, there is inevitably an unequal distance in the case of a broad spectrum, which cannot be corrected, but the unequal distance of the broadband interference has a small influence on the restoration result by controlling the thickness of the microsphere lens film 4. And controlling the diameter of the microsphere to be between 30 and 100 mu m by integrating the imaging field and the imaging quality, and the spin coating process and the immersion effect. The thickness of the thin film plate is between 50 and 200 μm in view of the spin coating process and the immersion effect.
In order to solve the problem of imaging quality reduction caused by spherical aberration, chromatic aberration and the like introduced by the microsphere lens film 4, the micro objective is optimized, and the objective, the light splitting flat plate, the reference flat plate, the microsphere flat plate and the film flat plate are combined to correct aberration in the range of the illumination light wide spectrum.
In the microsphere lens film 4, a film flat plate 6 is made of polydimethylsiloxane material, and a microsphere flat plate 7 is made of barium titanate material.
Fused quartz is used as glass materials of the reference flat plate 2 and the light splitting flat plate 3, and the thickness of the reference flat plate 2 and the thickness of the light splitting flat plate 3 are both 0.8-1.2 mm.
The diameter of the microspheres in the microsphere lens film 4 is between 30 and 100 micrometers, and the thickness of the film flat plate 6 is between 50 and 200 micrometers. Example one
In this example, we use a 50 μm-caliber barium titanate microsphere array combined with a PDMS film to make a microsphere lens film 4, and the specific preparation method is disclosed in patent 201610113464.2. The total thickness of the microsphere lens film 4 is 100 μm. And determining the thickness information of the reference flat plate 2 and the light splitting flat plate 3 according to an equation of the aplanatism at the central wavelength. The material fused quartz (refractive index 1.4606, Abbe number 67.82) of the light splitting plate 3 is 1mm in thickness, the material fused quartz (refractive index 1.4606, Abbe number 67.82) of the reference plate 2 is 1.132 mm in thickness, the distance between the reference plate 2 and the light splitting plate 3 is 3 mm, and the distance between the reference plate and the microsphere lens film is 3 mm.
In this example, the microsphere lens film 4 is equivalent to the superposition of the microsphere flat plate 6 and the film flat plate 7, the microsphere flat plate 5 is made of barium titanate (with the refractive index of 2.4109 and the Abbe number of 13.96) and has the thickness of 50 μm, and the film flat plate 7 is made of PDMS (with the refractive index of 1.4333 and the Abbe number of 44.44) and has the thickness of 50 μm. On the basis, the micro objective is optimized, and the objective, the light splitting flat plate, the reference flat plate and the microsphere lens film are combined to correct aberration in the range of the illumination light wide spectrum.
Taking a blue-ray disc, shearing partial area by using scissors, removing the protective film on the surface, and exposing parallel strips with the cycle of 200 nm and the interval of 100 nm to be used as a sample 5 to be detected.
The manufactured microsphere lens film 4 is placed on the surface of a sample 5 to be detected, and the optimized 50-time Mirau type interference microscope objective is carried on a white light microscope interferometer. By the adjustment, the interference fringes of the blu-ray disc are obtained as shown in fig. 3. From the interference fringes obtained in fig. 3, the three-dimensional topography of the blu-ray disc is obtained, as shown in fig. 4.
Claims (4)
1. A Mirau type super-resolution interference microscope objective is characterized in that: the micro-objective lens spectrometer comprises a micro-objective lens (1), a reference flat plate (2), a light splitting flat plate (3) and a micro-ball lens film (4) which are sequentially arranged along a common optical axis, wherein the micro-ball lens film (4) is arranged on the top surface of a sample (5) to be tested, a Mirau type interference optical path structure is adopted, light is split by the light splitting flat plate (3), the reference flat plate (2) is positioned on a reference arm, and the micro-ball lens film (4) is positioned on a test arm;
the illumination light (9) sequentially passes through the microscope objective (1) and the reference flat plate (2), is divided into two beams at the light splitting plane of the light splitting flat plate (3), one beam is incident to the reference surface of the reference flat plate (2) and returns to form reference light, and the other beam is incident to a sample to be tested (5) and returns after passing through the microsphere lens film (4) to form test light; the reference light and the test light are overlapped on the light splitting plate (3) and then interfere with each other, and the reference light and the test light are emitted out after passing through the reference plate (2) and the microscope objective (1);
at the center wavelength λ of the broad spectrum illumination light0The sum of the optical paths of the light splitting plate (3) and the microsphere lens film (4) is equal to the optical path of the reference plate (2);
the microsphere lens film (4) is prepared by coating a layer of film on the surface of a microsphere array, the microsphere array is equivalent to a microsphere flat plate (7), and the film is equivalent to a film flat plate (6); the refractive index of the film flat plate (6) is the refractive index of the film, and the thickness of the film flat plate (6) is the thickness of the microsphere lens film (4) minus the thickness of the microsphere flat plate (7); the refractive index of the microsphere flat plate (7) is the refractive index of the microsphere, and the thickness of the microsphere flat plate (7) is the diameter of the microsphere; at a central wavelength λ0Next, the following equation is satisfied:
n1t1+h1=n2t2+h2+n3t3+n4t4
wherein n is1For reference to the refractive index of the plate 1, n2Is the refractive index of the beam-splitting plate (2), t1For reference to the thickness of the plate 1, t2Is the thickness of the light-splitting plate (2), h1Is the spacing between the reference plate 1 and the spectroscopic plate 2, h2Is the interval between the light splitting plate (2) and the film plate (6), t3Is the thickness of the film plate (6), n3Is the refractive index, t, of the thin film plate (6)4Is the diameter of the microsphere flat plate (7), n4Is the refractive index of the microsphere flat plate (7);
fused quartz is used as glass materials of the reference flat plate (2) and the light splitting flat plate (3), and the thickness of the reference flat plate (2) and the thickness of the light splitting flat plate (3) are both 0.8-1.2 mm;
the diameter of the microspheres in the microsphere lens film (4) is 30-100 mu m, and the thickness of the film flat plate (6) is 50-200 mu m.
2. The Mirau-type super-resolution interference microscope objective lens according to claim 1, characterized in that: in the reference flat plate (2), the surface close to the microscope objective (1) is a reference surface; in the light-splitting flat plate (3), the surface close to the reference flat plate (2) is a light-splitting plane.
3. The Mirau-type super-resolution interference microscope objective lens according to claim 1, characterized in that: the magnification of the microscope objective (1) is not less than 10 times.
4. The Mirau-type super-resolution interference microscope objective lens according to claim 1, characterized in that: in the microsphere lens film (4), a film flat plate (6) is made of polydimethylsiloxane material, and a microsphere flat plate (7) is made of barium titanate material.
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| CN115755372A (en) * | 2022-11-12 | 2023-03-07 | 南京师范大学 | Transparent film applied to super-resolution imaging |
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| CN110376756A (en) * | 2019-07-10 | 2019-10-25 | 中国科学院光电技术研究所 | A kind of super-resolution microscopic system based on microballoon film |
| CN114923435B (en) * | 2022-04-29 | 2024-06-07 | 南京理工大学 | Cylindrical micro-interference device for measuring phase tomography information of micro-column optical components |
| CN115031623A (en) * | 2022-05-12 | 2022-09-09 | 中国电子科技集团公司第十一研究所 | A white light interference microscope objective |
| CN118705998A (en) * | 2024-07-15 | 2024-09-27 | 中国人民解放军国防科技大学 | Super-resolution three-dimensional shape measurement device and method combining microsphere lens and equivalent microsphere |
| CN119043195B (en) * | 2024-10-31 | 2025-04-29 | 贝耐特光学科技(苏州)有限公司 | Global detection device for LCoS liquid crystal cell thickness based on low-coherence light Mirau interferometry |
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